title: Robust ZFN-mediated genome editing in adult hemophilic mice.
abstract: Monogenic diseases, including hemophilia, represent ideal targets for genome-editing approaches aimed at correcting a defective gene. Here we report that systemic adeno-associated virus (AAV) vector delivery of zinc finger nucleases (ZFNs) and corrective donor template to the predominantly quiescent livers of adult mice enables production of high levels of human factor IX in a murine model of hemophilia B. Further, we show that off-target cleavage can be substantially reduced while maintaining robust editing by using obligate heterodimeric ZFNs engineered to minimize unwanted cleavage attributable to homodimerization of the ZFNs. These results broaden the therapeutic potential of AAV/ZFN-mediated genome editing in the liver and could expand this strategy to other nonreplicating cell types.
authors.affiliation: Division of Hematology and Center for Cellular and Molecular Therapeutics, Children's Hospital of Philadelphia, Philadelphia, PA;
authors.forename: Xavier M
authors.initials: XM
authors.lastname: Anguela
authors.forename: Rajiv
authors.initials: R
authors.lastname: Sharma
authors.forename: Yannick
authors.initials: Y
authors.lastname: Doyon
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Miller
authors.forename: Hojun
authors.initials: H
authors.lastname: Li
authors.forename: Virginia
authors.initials: V
authors.lastname: Haurigot
authors.forename: Michelle E
authors.initials: ME
authors.lastname: Rohde
authors.forename: Sunnie Y
authors.initials: SY
authors.lastname: Wong
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Davidson
authors.forename: Shangzhen
authors.initials: S
authors.lastname: Zhou
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.forename: Michael C
authors.initials: MC
authors.lastname: Holmes
authors.forename: Katherine A
authors.initials: KA
authors.lastname: High
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics; D004195:Disease Models, Animal; D004720:Endonucleases / Q000235:genetics* / Q000378:metabolism; D005164:Factor IX / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D016678:Genome; D002836:Hemophilia B / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D008099:Liver / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D055503:Protein Multimerization; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D005164:Factor IX; D004720:Endonucleases